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Aims-To determine the human papillomavirus DNA status of schistosomal associated squamous cell carcinoma of the urinary bladder in South Africa. Methods-Twenty five archival samples of bladder squamous cell carcinoma associated with Schistosoma haematobium were subjected to non-isotopic in situ hybridisation and the polymerase chain reaction for the detection of human papillomavirus 6,11,16,18, 31 Twenty five paraffin wax embedded tissue samples of bladder carcinomas from individuals infected with S haematobium were obtained from the departmental archives. All tumours were reviewed and classified as invasive squamous cell carcinomas, closely associated with ova of S haematobium. NON-ISOTOPIC IN SITU HYBRIDISATIONNon-isotopic in situ hybridisation (NISH) was performed using a technique described previously.6 7Sections (4 gm) from the paraffin wax embedded tissue samples were cut on to slides pretreated with aminopropyltriethoxysilane (Sigma, St Louis, Missouri, USA). The sections were allowed to dry overnight at 42°C and then dewaxed and rehydrated according to standard protocols. The slides were treated with 3% hydrogen peroxide in methanol to reduce residual non-specific peroxidase activity. Unmasking of nucleic acids was achieved by a limited proteolysis in proteinase K (500 jg/ml) at 37°C and the reaction stopped in distilled water after 15 minutes. The slides were air dried prior to the addition of aliquots of hybridisation mix (6 ,l) containing 2 ng/[tl of digoxigenin labelled HPV 6,11,16,18, 31, or

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